PhD Thesis
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Item Development of heteroploids in ginger (Zingiber off zingiber officinale Rosc.)(Department of Plantation Spices Medicinal and Aromatic Crops, College of Agriculture, Vellayani, 2024-01-24) Silpa, S G; Sreekala, G SAn experiment entitled “Development of heteroploids in ginger (Zingiber officinale Rosc.)” was carried out in the Department of Plantation, Spices, Medicinal and Aromatic Crops, College of Agriculture, Vellayani, during October 2018 to October 2022 with an objective to develop heteroploids using colchicine for yield and quality in ginger cultivars. The present study to develop heteroploids in ginger (Zingiber officinale Rosc.)” was carried out as three experiments (i) Development of heteroploids in ginger cultivars using colchicine in vivo (ii) Development of heteroploids in ginger cultivars using colchicine in vitro and (iii) Screening of heteroploids for stability. Experiments conducted at Department of Plantation, Spices, Medicinal and Aromatic Crops during 2018 recorded four promising genotypes of ginger. Rhizomes of these genotypes along with rhizomes of ginger varieties Athira, Aswathy, IISR Varada and IISR Mahima were collected from College of Agriculture, Vellanikkara and ICAR- Indian Institute of Spices Research respectively and heteroploidy was induced in vivo as per the method developed by Sheeba (1996). A total of sixteen treatments including eight respective controls were tried in Completely Randomized Design.Item Evaluation of CO2 enrichment on growth, development and soft rot tolerance in ginger (Zingiber officinale Rosc.)(Department of Plant Physiology, College of Agriculture, Vellayani, 2021) Manasa, R; Manju, R VThe present investigation entitled “Evaluation of CO 2 enrichment on growth, development and soft rot tolerance in ginger (Zingiber officinale Rosc.)” was undertaken with the objective to assess the impact of elevated CO 2 on growth, yield, quality and tolerance to soft rot in ginger through morphological, physiological, biochemical and molecular analysis. Two sets of pot culture experiments were conducted during 2017- 2019. The technology utilized for CO 2 enrichment was Open Top Chamber (OTC) system established under Department of Plant Physiology, College of Agriculture, Vellayani. In the first experiment three varieties of ginger Aswathy, Athira and Maran were utilized to assess the impact of elevated CO 2 on growth, yield and quality of ginger. One set of three month old plants were shifted to OTC and another set was retained under ambient CO 2. The plants were harvested at 4 th , 5 th , 7 th and 8 th months after planting (MAP). Observations on growth, physiological and biochemical parameters were recorded at each harvest. Yield, yield parameters, tissue nutrient status and quality parameters were assessed at 8 MAP. Second experiment was conducted to assess the impact of elevated CO 2 on tolerance to soft rot in three ginger varieties Aswathy, Athira, Maran and wild ginger congener Zingiber zerumbet. One set of three and a half month old plants were shifted to OTC and another set retained at ambient CO 2 and were challenged with Pythium aphanidermatum at 4 MAP. The CO 2 concentration of 500 ppm was maintained in the OTC. The results of the first experiment indicated an improvement in growth parameters like plant height, number of leaves, root weight, shoot weight, stem diameter and number of tillers, physiological and biochemical parameters like total chlorophyll, photosynthetic rate and total sugars in the rhizome by recording significantly higher values upon CO 2 enrichment when compared to ambient CO 2 condition. Significant reduction in transpiration rate, stomatal conductance and specific leaf area were recorded upon CO 2 enrichment. The per cent increase in yield parameters like number of fingers (38.19%), rhizome spread (11.32%), thickness (42.72%) and dry weight of rhizomes (31.43%) recordedhigher values upon CO 2 enrichment, based on average performance of all the selected varieties, leading to greater yield. The per cent increase in the fresh weight of rhizome recorded higher for variety Aswathy (38.34%), followed by Maran (20.00%) and Athira (12.12%) with a total fresh rhizome yield of 165.35 g plant -1 under ambient and 228.75 g plant -1 under elevated CO 2 condition in the case of Aswathy. Upon CO 2 enrichment the quality of rhizome was modified with increase in fiber (60.26%), gingerol (9.06%), phenol (19.25%), total oleoresin (11.52%), starch (44.84%) and essential oils (31.10%). Significant reduction in protein and nutritional status in the rhizomes were also recorded. Nitrogen, potassium, iron and zinc contents decreased by 15.78%, 3.22%, 11.78% and 18.57% respectively. Phosphorous and copper showed decrease in trend though not significant. Among the three varieties of ginger, Aswathy was found to be performing better compared to Athira and Maran not only in terms of yield, also for majority of quality parameters assessed upon exposure to elevated CO 2 condition. In the case of impact of elevated CO 2 on soft rot development, there was no variation in the per cent disease incidence. However, lesser number of days was taken for development of symptom. The per cent disease index was reduced by 11.10% upon CO 2 enrichment. The reduction in the fresh weight of rhizome upon pathogen inoculation was also less under elevated CO 2 (14.34%) when compared to ambient CO 2 (17.60%) showing that increasing CO 2 levels might bring down the disease severity. Upon P. aphanidermatum inoculation, the retention of photosynthetic pigments like chlorophyll a (0.90 mg g -1 FW) chlorophyll b (0.74 mg g -1 FW) total chlorophyll (1.64 mg g -1 FW) carotenoids (0.58 mg g -1 FW), xanthophyll (0.40 mg g -1 FW) and photosynthetic rate (5.92 μmol CO 2 m -2 s -1 ) were higher under elevated CO 2 when compared to ambient CO 2 . Higher membrane integrity values and lower malondialdehyde (0.75 μg g -1 FW) content in all the selected varieties were evident upon pathogen inoculation under CO 2 enrichment. Enhanced activity of ascorbate peroxidase (4.86 activity Units mg -1 total protein), PAL (Phenylalanine ammonia lyase) (20.91 Units mg -1 total protein), and glucanase (4.57Units mg -1 total protein) activity, as well as higher accumulation of phenols (32.40 mg g -1 DW) and ascorbic acid (7.14 mg 100 g -1 FW) were found in the inoculated plants under CO 2 enrichment compared to the ambient CO 2 . The catalase activity was found to reduce upon pathogen inoculation and the reduction was higher (0.11 Units mg -1 total protein) under ambient CO 2 as compared to elevated CO 2 (0.09 Units mg -1 total protein). Highest levels of hydrogen peroxide, superoxide and cell death were observed upon pathogen inoculation under ambient CO 2 through staining techniques and it was highest for variety Aswathy and least for Z. zerumbet. After pathogen inoculation maximum values for variable to maximum fluorescence Fv/Fm (0.63), photosynthetic electron transport rate (82.60 micro equiv m -2 s -1 ) and quantum yield of PS II photochemistry (0.29) were recorded under elevated CO 2. Protein profiling performed in varieties Athira and Aswathy revealed lower expressions of 20-25 kDa and 25-35 kDa protein under pathogen inoculation in variety Aswathy. The expression of 55 kDa (Rubisco Large Sub- Unit) was down regulated to a greater extent under pathogen inoculation in both the varieties. Higher expression levels of the defense responsive genes Phenylalanine ammonium-lyase (PAL), Cinnamyl alcohol dehydrogenase (CAD) and Chalcone synthase (CSH) were also observed upon pathogen inoculation under CO 2 enrichment in both the varieties. Between the varieties, Athira had higher relative fold changes of PAL, CAD and CSH under both ambient and elevated CO 2 upon pathogen inoculation. The present study revealed that CO 2 enrichment can improve the performance of ginger in terms of growth, yield and quality. It also indicates changes in photosynthetic allocation pattern and phytochemical profiles with enhanced contents of oleoresins and essential oils. Ginger plants showed better tolerance against P. aphanidermatum upon CO 2 enrichment. This was apparent in the form of reduced disease severity, oxidative stress level and less reduction in yield upon pathogen infestation under elevated CO 2 condition. This was evidently achieved through the activation of multiple mechanisms, including enhanced production of anti-oxidants and up regulation of pathogen resistance proteins. This is one of the first studies to comprehend on possible effects of elevated CO 2 on the susceptibility of ginger to soft rot. The study also demonstrated the varietal variations existing in the response to CO 2 enrichment which will help in selection ofsuitable varieties for changing climatic scenario. The information generated indicates the possibility of enhancing both qualitative and quantitative yield of ginger through low cost CO 2 enrichment technologies. The insights obtained on the tolerance mechanisms involved will help to restructure the soft rot disease management strategies in the coming future.Item In vitro synthesis of Gingerol and analysis of expressed sequence tags for Gingerol production in Ginger(Zingiber officinale Rosc.)(Centre for plant biotechnology and molecular biology,College of Horticulture, Vellanikkara, 2020) Manjusha, Rani; Shylaja, M RItem Effectiveness of soil solarization for the control of soft rot disease in ginger(Department of Plant Pathology, College of Horticulture, Vellanikkara, 1996) Vilasini, T N; Peethambran, C KItem Induction of variation in vitro and field evaluation of somaclones in ginger (zingiber officinale rosc)(Department of Plantation Crops and Spices, College of Horticulture, Vellanikkara, 2006) Resmi Paul; Shylaja, M RItem Agrobacterium mediated genetic transformation of ginger (zinbiger officinale rosc.)(Department of Plantation Crops and Spices, College of Horticulture, Vellanikkara, 2006) Suma, B; Keshavachandran, RInvestigations on genetic transformation in ginger (Zingiber officinale Rosc.) variety Rio-de-Janeiro using Agrobacterium tumefaciens strain EHA 105 harbouring antibiotic resistant selectable marker genes (npt II) and GUS reporter genes were carried out at the Department of Plantation Crops and Spices and Plant Tissue Culture Laboratory, CPBMB, College of Horticulture, Vellanikkara, Thrissur during the period from 1999 to 2005. Axenic shoot bud cultures of ginger variety Rio-de-Janeiro was raised under in vitro condition to generate explants with reduced contamination for transformation. Half strength MS medium with BA 3 mg l-1 was found to be the best for establishing shoot bud cultures. In order to standardise a regeneration protocol, MS medium supplemented with varying concentration of auxin and cytokinin were tried on different explants. Embryogenic calli were induced from bud explants of ginger supplemented with MS + 1.0 mg l-1 2,4-D + 0.5 mg l-1 BA, followed by plant regeneration on MS medium + BA 3 mg l-1 + 2,4-D 0.5 mg l-1. Bactericidal effect of antibiotics towards different strains of Agrobacterium and sensitivity of ginger tissues to different antibiotics were also studied to standardise the optimum level of antibiotics. Cefotaxime at a concentration of 300 mg l-1 was selected for eliminating the bacteria after co-cultivation. Kanamycin 100 mg l-1 was used to discriminate between transformed and untransformed cells. Agrobacterium strains were collected, recombinants were made and the presence of the construct confirmed in the native strains of Agrobacterium before starting transformation experiments. Agrobacterium strain EHA 105 p35SGUSINT was used for standardising the optimum conditions by comparing the levels of transient GUS expression in inoculated buds. A suitable transformation protocol would include 3 days preculture of explants, bacterial dilution of 1:20 (v/v), infection time of 5 min, co-cultivation of 48 h and post cultivation on callus induction medium with 100 mg l-1 kanamycin + 300 mg l-1 cefotaxime in darkness for 2 weeks and then under 16/8 h photoperiod. Use of acetosyringone in the co-cultivation medium (200 µm) and vir induced Agrobacterium strain (200 µm), increased the efficiency of transformation. Histochemical GUS assays were employed to study and compare the transient GUS expression, stable expression from putative transgenics. Further confirmation was made by PCR assays. The regeneration protocol as well as transformation protocol could be effectively used for further transformation.Item Production dynamics of ginger (zingiber officinale R.) under varying levels of shade, nutrients and triazole(Department of Horticulture, College of Agriculture, Vellayani, 1999) Ajith Kumar, K; Jayachandran, B KThe experiments were conducted at the Coconut Research Station, Balaramapuram, during 1996-97 and 1997-98 to study the production dynamics of ginger under varying levels of shade, nutrients and triazole. The specific objectives were to study the production potential of ginger under open and different shade levels, to standardise optimum dose of nutrients and mulch for ginger intercropped in coconut garden and to evaluate the efficacy of triazole in the improvement of yield and other desirable characters. The effect of shade on growth and yield contributing parameters viz., tiller number, leaf number, leaf area, leaf dry weight, root spread, root volume, rhizome spread, rhizome thickness, number of rhizomes, leaf thickness, stomatal frequency, DMP and LAI under 20 and 40 per cent shade levels exhibited significant superiority in pot culture study. In pot culture study: maximum fresh ginger yield of 450.0 and 396.3 g plant" were resulted from plants kept under 20 and 40 per cent shade levels respectively. This was 27.4 and 12.2 per cent higher compared to open- condition. The dry ginger yield of 94.5 and 89.2 g plant" were obtained from plants kept under 20 and 40 per cent shade levels respectively. This was 27.8 and 20.7 per cent higher compared to open condition. However, the dry ginger yield obtained from 60 and 80 per cent shade levels were significantly lower compared to open. The shade levels 20 and 40 per cent gave the highest shoot dry weight of 48.9 and 53.6 g plant" respectively. Volatile oil content showed an increasing trend with increasing levels of shade in pot culture study. Maximum NVEE was recorded under 20 and 40 per cent shade levels. Highest starch content was obtained from 20 per cent shade level. Crude fibre gradually reduced with increase in shade levels. The field trial on nutrient requirement of ginger as intercrop in coconut garden revealed that, NPK application at higher levels increased plant height, tillering, leaf number, leaf area, DMP and chlorophyll content. Physiological parameters like, NAR, CGR and RGR were promoted by higher rates of nitrogen application. The results revealed that, N: P, N : K and P : K ratios of l.5 : 1, l.5 : 1 and 1 : 1 respectively, were ideal to get maximum fresh ginger yield. Application of 150 kg N, 100 kg P20S and 100 kg K20 ha-l gave maximum net profit ofRs. 96605/- and Rs. 99227/- during first and second year respectively. The net profit when the existing recommendation of 75 kg N, 50 kg P20S and 50 kg K20 ha-l was Rs. 54960/- and Rs. 54730/- during first and second year respectively. Hence the study suggests that the existing recommendation (75 kg N, 50 kg P20S and 50 kg K20 ha") has to be doubled (150 kg N, 100 kg P20S and 100 kg K20 ha-l) for increasing the productivity of ginger intercropped in coconut garden. Increased rate of fertilizer application did not influence the quality of the produce. Plant uptake of NPK increased with higher levels of fertilizer application. The field experiment on mulch requirement of ginger intercropped in coconut garden showed that the rhizome yield from plots mulched with 30.0, 22.5 and 1 =r. 0 t ha' were on par. Therefore the mulch requirement of ginger intercropped in coconut garden can be reduced from 30 t ha-l to 15.0 t ha-I Application of triazole did not exhibit any beneficial effect on yield and quality of ginger.Item Optimisation of process parameters for preparation of flavoured instant green tea(Department of Food and Agricultural Process Engineering, Kelappaji College of Agricultural Engineering and Technology, Tavanur, 2017) Sankalpa, K B; Santhi Mary, MathewItem Metabolite Profiling and gene expression analysis for gingerol production in selected somaclones of ginger (zingiber officinale rosc.)(Centre for Plant Biotechnology and Molecular Biology, College of Horticulture, Vellanikkara, 2017) Sreeja, S; Shylaja, M RItem Screening ginger (Zingiber officinale Rosc.) genotypes under different growing conditions and for value addition(Department of Plantation Crops and Spices College of Horticulture, Vellanikkara, 2018) Nimisha Mathews; Krishnakumary, KGinger is an important commercial spice crop grown in India for culinary and medicinal purposes. The present study entitled “Screening ginger (Zingiber officinale Rosc.) genotypes under different growing conditions and for value addition” was taken up at College of Horticulture, Vellanikkara during 2015-18 to evaluate the performance of ginger genotypes with respect to yield and quality, growth under different growing conditions, quality of different maturity stages and to identify types suitable for different value added products. A total of fourteen somaclones developed through indirect organogenesis and indirect embryogenesis were selected for the study. Genotypes exhibited wide variability in plant characters, yield characters and quality attributes. Plant height was maximum in the somaclone SE 86102 (107.35 cm) at 6 months growth stage. Number of tillers was maximum in CHP118 (20.33) at 6 months stage. Number of leaves per shoot was found higher in CHP 118 (28.67) and in SE 86 26. Leaf area was the highest in CHP 118 at 4 months stage and C8632 at 6 months stage. Among the physiological parameters recorded, photosynthetic and transpiration rates were the highest in CHP 118 and SE 86 42. Number of primary and secondary rhizomes was maximum in SE 8626, CHP 118 and C8632 (>30 t/ha). Weight of primary and secondary rhizomes was highest in SE 8626 and SE 8642. Fresh yield was highest in SE 8626, CHP 118, SE 8642 & C 8632. Driage (23%) and dry yield (7.9t/ha) were the highest in SE 8626. The quality attributes such as volatile oil, oleoresin and crude fiber contents varied significantly among the genotypes. The highest content of volatile oil (3.62%) was recorded in CHP 118. Oleoresin content (>6%) was maximum in Rio-de- Jenairo. Fibre content increased with age of rhizomes recording lowest values (< 2.5%) in SE 86 83, C 86 26 and SE 86 42. Highest content of Gingerol (1.48 %) and Shogoal (0.16%) was recorded in Rio –de- Janeiro and CHP 282 respectively. Among the genotypes evaluated under different growing conditions, variability in plant height was observed. During the growth stages, all the genotypes exhibited higher plant height under rain shelter condition, whereas at 6 months stage plants were taller in open field. Number of tillers and leaves were significantly higher in open condition. Tiller number ranged from 14 to 17 under open field and 6 to 7 in rain shelter. Leaf area was the highest at 5 months growth stage which later decreased. Among the two growing conditions, the highest leaf area was observed in open condition in SE 8640 and SEHP 9. Generally, somaclones grown in open condition recorded higher yield and SEHP 9, SE 8081 and SE 8640 were identified as higher yielders. But in SE 86 40, the yield was on par both in open and rain shelter conditions indicating its suitability for growing under rain shelter condition. Driage did not differ significantly between the two growing conditions. Driage and dry yield were higher in SE 86 81 and SEHP 9. Higher values for photosynthetic rate, transpiration rate and stomatal conductance were recorded in open field. Among the two growing conditions, quality parameters were higher in rainshelter cultivation though not significant. SE 8640 recorded the highest volatile oil content under both growing conditions (open field -4.45 % and rain shelter -4.67 %). All genotypes except SE 8640 recorded an oleoresin content of more than 5 %. Fibre content increased with maturity recording maximum values at 7 months stage. Lower fibre content was recorded under rain shelter condition. Among the somaclones, SE 8681 and SE 86131 recorded lower values for fiber content at 5 months stage. Elite varieties satisfying the requirements for specific end products are the need of the hour to capitalize on the processing front. Biochemical parameters such as moisture content, TSS, titratable acidity, pH and colour were recorded for analyzing the storage stability of the products viz., candy, flakes and powder. The somaclones SE 8683, C 8626 and SE 8642 were found to be the best for candy preparation based on its overall consumer acceptability and storage stability. SE 8683, C 8626 and SE 8642 were found suitable for flakes preparation and SE 8683, C 8626, SE 8642 and C8632 for making quality ginger powder.